Functional Characterization and Localization of Acetyl-Coa Hydrolase, Ach 1p, in Saccharomyces Cerevisiae
Resource
JOURNAL OF BIOLOGICAL CHEMISTRY v.278 n.19 pp.17203-17209
Journal
JOURNAL OF BIOLOGICAL CHEMISTRY
Journal Volume
v.278
Journal Issue
n.19
Pages
17203-17209
Date Issued
2003
Date
2003
Author(s)
CHEN, YEE-CHUN
Abstract
Acetyl-CoA hydrolase (Ach1p), catalyzing the hydrolysis of acetyl-CoA, is presumably involved in regulating intracellular acetyl-CoA or CoASH pools ; however, its intracellular functions and distribution remain to be established. Using site-directed mutagenesis analysis, we demonstrated that the enzymatic activity of Ach1p is dependent upon its putative acetyl -CoA binding sites. The ach1 mutant causes a growth defect in acetate but not in other non-fermentable carbon sources, suggesting that Ach1p is not involved in mitochondrial biogenesis. Overexpression of Ach1p, but not constructs containing acetyl-CoA binding site mutations, in ach1-1 complemented the defect of acetate utilization. By subcellular fractionation, most of the Ach1 p in yeast was distributed with mitochondria and little Ach 1 p in the cytoplasm. By immunofluorescence microscopy, we show that Ach1p and acetyl-CoA binding site-mutated constructs, but not its N-terminal deleted construct, are localized in mitochondria. Moreover, the onset of pseudohyphal development in homozygote ach1-1 diploids was abolished. We infer that Ach1p may be involved in a novel acetyl-CoA biogenesis and/or acetate utilization in mitochondria and thereby indirectly affect pseudohyphal development in yeast.
Subjects
COENZYME-A HYDROLASE
AMINO-ACID-SEQUENCE
RAT-LIVER
ACETATE UTILIZATION
FILAMENTOUS GROWTH
PROTEIN IMPORT
Type
journal article